Phosphorylation of Rap1GAP, a striatally enriched protein, by protein kinase A controls Rap1 activity and dendritic spine morphology

Protein kinase A (PKA)-dependent signaling cascades play an important role in mediating the effects of dopamine and other neurotransmitters in striatal medium spiny neurons. We have identified a prominent striatal PKA substrate as Rap1-GTPase activating protein (Rap1GAP), a negative regulator of Rap1 signaling. Although present throughout the brain, Rap1GAP is enriched in striatal medium spiny neurons and is phosphorylated by PKA at Ser-441 and Ser-499 in response to activation of D1 dopamine receptors. Phosphorylation of Rap1GAP is associated with inhibition of GAP activity, as demonstrated by increased Rap1 activity in striatal neurons. Phosphorylation of Rap1GAP is also associated with increased dendritic spine head size in cultured neurons. These findings suggest that phosphorylation of Rap1GAP by PKA plays an important role in striatal neurons by modulating Rap1 actions.

[1]  F. McCormick,et al.  Localization of the rap1GAP catalytic domain and sites of phosphorylation by mutational analysis , 1992, Molecular and cellular biology.

[2]  W. Kabsch,et al.  The Ras-RasGAP complex: structural basis for GTPase activation and its loss in oncogenic Ras mutants. , 1997, Science.

[3]  A. Wittinghofer,et al.  Epac is a Rap1 guanine-nucleotide-exchange factor directly activated by cyclic AMP , 1998, Nature.

[4]  P. Greengard,et al.  Ca2+/calmodulin-dependent protein kinase II: identification of autophosphorylation sites responsible for generation of Ca2+/calmodulin-independence. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Xiaodong Cheng,et al.  Cyclic adenosine 5'-monophosphate-stimulated neurotensin secretion is mediated through Rap1 downstream of both Epac and protein kinase A signaling pathways. , 2007, Molecular endocrinology.

[6]  P. Greengard,et al.  Beyond the Dopamine Receptor: Review the DARPP-32/Protein Phosphatase-1 Cascade , 1999 .

[7]  T. Kataoka,et al.  Effect of Phosphorylation on Activities of Rap1A to Interact with Raf-1 and to Suppress Ras-dependent Raf-1 Activation* , 1999, The Journal of Biological Chemistry.

[8]  J. Bos,et al.  Epac proteins: multi-purpose cAMP targets. , 2006, Trends in biochemical sciences.

[9]  Richard L. Huganir,et al.  Activity-Dependent Dendritic Spine Structural Plasticity Is Regulated by Small GTPase Rap1 and Its Target AF-6 , 2005, Neuron.

[10]  Ravi Iyengar,et al.  Modulation of Rap Activity by Direct Interaction of Gαo with Rap1 GTPase-activating Protein* , 1999, The Journal of Biological Chemistry.

[11]  P. Greengard,et al.  A Network of Control Mediated by Regulator of Calcium/Calmodulin-Dependent Signaling , 2004, Science.

[12]  J. Bos,et al.  Ras and Rap1: two highly related small GTPases with distinct function. , 1999, Experimental cell research.

[13]  Paul Greengard,et al.  Cell type–specific regulation of DARPP-32 phosphorylation by psychostimulant and antipsychotic drugs , 2008, Nature Neuroscience.

[14]  M. Harbeck,et al.  Cell physiology of cAMP sensor Epac , 2006, The Journal of physiology.

[15]  J. Girault,et al.  Opposing Patterns of Signaling Activation in Dopamine D1 and D2 Receptor-Expressing Striatal Neurons in Response to Cocaine and Haloperidol , 2008, The Journal of Neuroscience.

[16]  P. Stork,et al.  Multiple roles of Rap1 in hematopoietic cells: complementary versus antagonistic functions. , 2005, Blood.

[17]  Angus C Nairn,et al.  Cocaine-induced dendritic spine formation in D1 and D2 dopamine receptor-containing medium spiny neurons in nucleus accumbens. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[18]  A. Wittinghofer,et al.  Rap-specific GTPase Activating Protein follows an Alternative Mechanism* , 2002, The Journal of Biological Chemistry.

[19]  Anatol C. Kreitzer,et al.  Striatal Plasticity and Basal Ganglia Circuit Function , 2008, Neuron.

[20]  Bryan Kolb,et al.  Structural plasticity associated with exposure to drugs of abuse , 2004, Neuropharmacology.

[21]  N. Minato,et al.  Human SPA-1 Gene Product Selectively Expressed in Lymphoid Tissues Is a Specific GTPase-activating Protein for Rap1 and Rap2 , 1997, The Journal of Biological Chemistry.

[22]  D. Surmeier,et al.  D1 and D2 dopamine-receptor modulation of striatal glutamatergic signaling in striatal medium spiny neurons , 2007, Trends in Neurosciences.

[23]  I. Medina.  Extrasynaptic NMDA Receptors Reshape Gene Ranks , 2007, Science's STKE.

[24]  Paul Greengard,et al.  DARPP-32: Regulator of the Efficacy of Dopaminergic Neurotransmission , 1998 .

[25]  M. Sheng,et al.  Regulation of Dendritic Spine Morphology by SPAR, a PSD-95-Associated RapGAP , 2001, Neuron.

[26]  R. Malinow,et al.  Ras and Rap Control AMPA Receptor Trafficking during Synaptic Plasticity , 2002, Cell.

[27]  P. Casey,et al.  Activation of Gz Attenuates Rap1-mediated Differentiation of PC12 Cells* , 2002, The Journal of Biological Chemistry.

[28]  A. Sizeland,et al.  The residues of Ras and Rap proteins that determine their GAP specificities. , 1991, The Journal of biological chemistry.

[29]  Paolo Calabresi,et al.  Dopamine-mediated regulation of corticostriatal synaptic plasticity , 2007, Trends in Neurosciences.

[30]  P. Greengard,et al.  Localization of ARPP-90, a major 90 kiloDalton basal ganglion-enriched substrate for cyclic AMP-dependent protein kinase, in striatonigral neurons in the rat brain. , 1989, Brain research. Molecular brain research.

[31]  J. Meinkoth,et al.  Thyroid-stimulating Hormone/cAMP and Glycogen Synthase Kinase 3β Elicit Opposing Effects on Rap1GAP Stability* , 2004, Journal of Biological Chemistry.

[32]  P. Greengard,et al.  A Translational Profiling Approach for the Molecular Characterization of CNS Cell Types , 2008, Cell.

[33]  P. Greengard,et al.  DARPP-32, a dopamine- and adenosine 3':5'-monophosphate-regulated phosphoprotein enriched in dopamine-innervated brain regions. I. Regional and cellular distribution in the rat brain , 1984, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[34]  H. Yao,et al.  cAMP Activates MAP Kinase and Elk-1 through a B-Raf- and Rap1-Dependent Pathway , 1997, Cell.

[35]  P. Stork,et al.  PKA phosphorylation of Src mediates Rap1 activation in NGF and cAMP signaling in PC12 cells , 2004, Journal of Cell Science.

[36]  P. Greengard.  The neurobiology of slow synaptic transmission. , 2001, Science.

[37]  E. Kandel,et al.  Rap1 Couples cAMP Signaling to a Distinct Pool of p42/44MAPK Regulating Excitability, Synaptic Plasticity, Learning, and Memory , 2003, Neuron.

[38]  J. Bos,et al.  Rap1: a key regulator in cell-cell junction formation , 2006, Journal of Cell Science.

[39]  Angus C Nairn,et al.  Protein kinase A activates protein phosphatase 2A by phosphorylation of the B56δ subunit , 2007, Proceedings of the National Academy of Sciences.

[40]  E. Lapetina,et al.  Mutational analysis of the cAMP-dependent protein kinase-mediated phosphorylation site of Rap1b. , 1993, The Journal of biological chemistry.

[41]  M. Avolio,et al.  Rap1: a turnabout for the crosstalk between cadherins and integrins. , 2006, European journal of cell biology.

[42]  P. Greengard,et al.  ARPP-21, a cyclic AMP-regulated phosphoprotein enriched in dopamine- innervated brain regions. I. Purification and characterization of the protein from bovine caudate nucleus , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[43]  L. Birnbaumer,et al.  Compartmentalization of protein kinase A signaling by the heterotrimeric G protein Go , 2006, Proceedings of the National Academy of Sciences.

[44]  A. Graybiel,et al.  A Rap guanine nucleotide exchange factor enriched highly in the basal ganglia. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[45]  P. Greengard,et al.  Regional distribution of calcium- and cyclic adenosine 3':5'- monophosphate-regulated protein phosphorylation systems in mammalian brain. II. Soluble systems , 1983, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[46]  J. Bos.  Linking Rap to cell adhesion. , 2005, Current opinion in cell biology.

[47]  S. Døskeland,et al.  Cyclic adenosine 3',5'-monophosphate (cAMP)-dependent protein kinases, but not exchange proteins directly activated by cAMP (Epac), mediate thyrotropin/cAMP-dependent regulation of thyroid cells. , 2007, Endocrinology.

[48]  P. Greengard,et al.  Dichotomous Dopaminergic Control of Striatal Synaptic Plasticity , 2008, Science.

[49]  P. Greengard,et al.  DARPP-32, a dopamine- and adenosine 3':5'-monophosphate-regulated phosphoprotein enriched in dopamine-innervated brain regions. III. Immunocytochemical localization , 1984, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[50]  J. Bos,et al.  Activation of the small GTPase rap1 in human neutrophils. , 1998, Blood.

[51]  V. Gallo,et al.  cAMP-dependent Protein Kinase Induces cAMP-response Element-binding Protein Phosphorylation via an Intracellular Calcium Release/ERK-dependent Pathway in Striatal Neurons* , 2001, The Journal of Biological Chemistry.

[52]  Paul Greengard,et al.  A dopamine- and cyclic AMP-regulated phosphoprotein enriched in dopamine-innervated brain regions , 1983, Nature.

[53]  F. McCormick,et al.  Phosphorylation of rap1GAP in vivo and by cAMP-dependent kinase and the cell cycle p34cdc2 kinase in vitro. , 1992, The Journal of biological chemistry.

[54]  F. McCormick,et al.  Molecular cloning of a GTPase activating protein specific for the Krev-1 protein p21 rap1 , 1991, Cell.